ObjectiveTo study the characteristics of imprinting template of flavonoid clusters in four Chinese medicines attributed to the lung meridian， and to establish an in vitro experimental approach for the study of the attribution of Chinese medicines to the lung meridian. MethodBased on 13 Chinese medicines， including Xanthii Fructus， Houttuyniae Herba， Fagopyri Dibotryis Rhizoma， Belamcandae Rhizoma and so on， which only belong to the lung meridian in Chinese Materia Medica（the 13^th Five-Year planning textbook of general higher education）， we identified four representative Chinese medicines， namely Houttuyniae Herba， Fagopyri Dibotryis Rhizoma， Belamcandae Rhizoma and Mori Cortex， and set up their fingerprints by high performance liquid chromatography（HPLC） and calculated the molecular connectivity indices of various components in the four Chinese medicines， the similarity to their mean value was calculated by included angle cosine method， so as to establish the quantitative relationship of construction versus imprinting ability， and to determine the order of each component in the lung meridian. A total of 7 reference substances， including chlorogenic acid， rutin， quercetin， isoquercitrin， hyperoside， epicatechin， and iridin， were selected to validate the overall conformational relationships of flavonoids of the model， as well as its predictive ability. ResultHouttuyniae Herba， Fagopyri Dibotryis Rhizoma， Belamcandae Rhizoma and Mori Cortex contained a total of 437 chemical components with an average molecular connectivity index similarity of 0.995 6. The four Chinese medicines contained a total of 204 flavonoids with an average molecular connectivity index similarity of 0.978 0， which was second only to the alkaloids with 0.985 1. The retention time（t_R） of the 7 reference substances showed a good conformational relationship with the similarity of the molecular connectivity index（t_R=831.4×S-790.3， r=0.861 4， P<0.01）， which was applicable to the in vitro attribution study of the position， similarity， and relative similarity with t_R of the cluster of 98.04% of flavonoids. Accordingly， the 1^st position was kuwanon D， with a similarity of molecular connectivity index of 0.987 7 and a t_R of 30.88 min， the 200^th position was chlorogenic acid， with a similarity of molecular connectivity index of 0.958 2 and a t_R of 6.36 min. The total first-order moment of the four Chinese medicines calculated by total statistical moment method of fingerprint was 24.26 min， ranked 21， which could characterize 99.19% of the whole， and the total first-order moment of the total peak area of the 7 reference substances in the four Chinese medicines was 20.00 min， with a rank of 46， which could characterize 98.68% of the whole. ConclusionFlavonoid clusters are suitable probes for the characterization of imprinting template for the study of the lung meridian， which can be established a quantitative imprinting method for meridian tropism of Chinese medicines in vitro.

Objective To explore the clinical efficacy of Fuling Zexie Decoction combined with lifestyle intervention in patients with metabolic syndrome (MetS) of dampness syndrome. Methods A total of 40 patients with MetS of dampness syndrome were randomly divided into experimental group (n=20) and control group (n=20) according to a randomized double-blind placebo-controlled design. The experimental group was given oral administration of Fuling Zexie Decoction,the control group was given placebo,150 mL each time,twice a day. Both groups were given healthy lifestyle(healthy eating habits and exercise mode) education. The intervention period was 16 weeks. Traditional Chinese medicine dampness syndrome scale score and the diagnostic indexes of MetS,including waist circumference (WC),systolic blood pressure (SBP),diastolic blood pressure (DBP),triglyceride (TG),high density lipoprotein cholesterol (HDL-C) and fasting blood glucose (FBG),were evaluated before and after treatment. Other metabolic indexes such as body mass index(BMI),total cholesterol(TC),low density lipoprotein cholesterol(LDL-C),homeostasis model assessment of insulin resistance(HOMA-IR),uric acid(UC),as well as inflammation related indexes including C-reactive protein (CRP),interleukin-6 (IL-6) and tumor necrosis factor(TNF-α) were also evaluated. Results A total of 33 people completed the trial,including 17 people in the experimental group and 16 people in the control group. Compared to the baseline period,FBG,BMI,LDL-C and dampness syndrome scale score in the experimental group were significantly decreased at the 8th and 16th week of intervention(P<0.05). WC was significantly decreased at the 8th week of intervention(P<0.05). CRP showed a downward trend at the 16th week of intervention(P<0.05). Moreover,SBP and IL-6 in control group decreased significantly at the 8th and 16th week of intervention(P<0.05). WC,DBP and HOMA-IR decreased significantly at the 8th week of intervention (P<0.05). There was no significant difference in other outcome indexes between the two groups at the 8th and 16th week of intervention (P>0.05). The safety indexes of all patients before and after intervention were not significantly abnormal,and no serious adverse events occurred. Conclusion This study initially shows the positive regulatory effect of Fuling Zexie Decoction combined with lifestyle on metabolism and inflammation-related indexes in people with MetS of dampness syndrome,which helps to improve the level of dampness syndrome and provides a basis for further research.

Objective To explore the relationship of UHRF1 with the clinicopathological characteristics of colorectal cancer (CRC) patients, as well as the effects of lentivirus transfection overexpression and knockdown of UHRF1 on the proliferation, invasion, and migration of CRC cells and the possible signaling pathways. Methods The expression of UHRF1 mRNA and protein in CRC tissues and adjacent tissues was detected by immunohistochemical staining and RT-PCR. The effects of the constructed UHRF1 overexpression- and knockdown-group cells on the expression of UHRF1, related molecules in the WNT signaling pathway, and MMPR9 were examined by Western blot and RT-PCR. EDU and Transwell assays were used to detect changes in the proliferation, migration, and invasion of CRC cells. Results (1) In the TCGA database and clinical data, the mRNA and protein expression levels of UHRF1 in CRC cancer tissues were significantly higher than those in adjacent normal tissues. UHRF1 expression was closely correlated with TNM stage, N stage, and M stage. Patients with low UHRF1 expression in TCGA had better 5-year OS and disease-specific survival. The area under the ROC curve of UHRF1 for predicting 1-, 3-, and 5-year OS were 0.634, 0.652, and 0.771, respectively. The 3-year OS in the clinical data also showed the same survival benefit. UHRF1 overexpression was a poor prognostic factor for CRC patients. (2) After UHRF1 overexpression, the expression of WNT3a, GSK3β, and MMP9 in SW480 cells significantly increased, whereas the expression of p-β-catenin decreased (P < 0.05). After UHRF1 knockdown, the expression of WNT3a, GSK3β, and MMP9 in HCT116 cells decreased, whereas the expression of p-β-catenin increased (P < 0.05). The "rescue" experiment with IWP-2 and HLY78 can produce consistent results. (3) Compared with the control group, the cell proliferation, migration, and invasion abilities of the UHRF1 overexpression group were enhanced. After IWP-2 treatment, the cell proliferation, migration, and invasion abilities were inhibited. Knockdown experiment exhibited the reverse results to overexpression experiment. Conclusion UHRF1 may play an important role in the occurrence and development of CRC. UHRF1 overexpression may be a poor prognostic factor for CRC patients. UHRF1 may affect the proliferation, migration, and invasion of CRC cells through the WNT/MMP9 signaling pathway.

Objective:To observe the efficacy and safety of teriprizumab combined with bevacizumab in above the second line treatment of high-level microsatellite instability (MSI-H) type metastatic colorectal cancer (mCRC) patients.Methods:From February 2019 to September 2019, 56 patients with MSI-H mCRC admitted to the Third Affiliated Hospital of Guangdong Medical University were selected and divided into control group and test group by random number table method, with 28 cases in each group. The control group was treated with bevacizumab, and the test group was treated with teriprizumab combined with bevacizumab. The objective response rate (ORR), disease control rate (DCR), progression-free survival, overall survival and incidence of adverse reactions were compared between the two groups.Results:The ORR and DCR of the test group were 60.71% (17/28) and 75.00% (21/28) respectively, higher than 28.57% (8/28) and 46.63% (13/28) of the control group, with statistically significant differences ( χ2=5.85, P=0.016; χ2=4.79, P=0.029). The median progression-free survival of patients in the control group and the test group were 3.5 months and 5.8 months respectively, with a statistically significant difference ( χ2=9.83, P=0.003). The median overall survival of patients in the control group and the test group were 12.1 months and 16.2 months respectively, with a statistically significant difference ( χ2=6.13, P=0.007). There were no significant diffe-rences in the incidences of hematological reaction (17.86% vs. 14.29%, χ2=0.13, P=0.716), cardiovascular injury (10.71% vs. 14.29%, χ2=0.16, P=0.686), liver and kidney function injury (25.00% vs. 21.43%, χ2=0.10, P=0.752), gastrointestinal reaction (28.57% vs. 35.71%, χ2=0.33, P=0.567), skin and mucosal injury (7.14% vs. 10.71%, χ2=0.35, P=0.553), nervous system disease (3.57% vs. 14.29%, χ2=2.25, P=0.134), endocrine reaction (3.57% vs. 10.71%, χ2=1.29, P=0.256), alopecia (14.29% vs. 17.86%, χ2=0.13, P=0.716) and fatigue (25.00% vs. 28.57%, χ2=0.27, P=0.605) between the control group and the test group. Conclusion:The combination of teriprizumab and bevacizumab can improve the short-term and medium-long-term efficacy of patients with MSI-H mCRC, which is safe and reliable.

Objective:To explore the effects of KISS1 gene transfected by lentivirus on the proliferation,invasion and migration abilities of the colorectal cancer HCT116 cells,and to clarify their mechanisms.Methods:The human colorectal cancer cells with the lowest expression level of KISS1 gene were selected.The lentiviral vectors were builted and transfected the KISS1 gene,and the cells were divided into control group (treated with PBS),empty vector group (treated with empty vector) and over-expression group(treated with KISS1 gene vector).The multiplicity of infection (MOI) of the cells was detected by fluorescence microscope.Real-time PCR and Western blotting methods were used to detect the expression levels of KISS1 mRNA and protein(metastin);CCK-8 method was used to detect the proliferation ability of the cells;Transwell chambers method was used to detect the invasion and migration abilities of the cells.Results:Among LoVo,SW620,SW480,HCT-116,and HT29 cells,the expression levels of KISS1 mRNA and protein were lowest in HCT116 cells,so they were chosen as the research carrier.After transfected with lentiviral vectors,the HCT116 cells could stably express the enhanced green fluorescent protein(EGFP) gene,and the MOI was over 80%.Compared with control group and empty vector group,the expression levels of KISS1 mRNA and protein in the cells in over-expression group were significantly increased (P<0.05);the proliferation abilities of the cells in over-expression group were decreased (P<0.05);the invasion ability and migration ability of the cells in over-expression group were decreased (P<0.05).But the differences of proliferation ability,invasion ability and migration ability of the cells between control group and empty vector group were not statistically significant (P>0.05).Conclusion:The KISS1 gene transfected by lentivirus vector can over-express KISS1 protein and inhibit the proliferation,invasion and migration abilities of the colorectal cancer cells,and the mechanism may be related to the expression of KISS1 protein.

Objective To study clinical effect after laparoscopic abdominal preperitoneal inguinal hernia repair methodwithout stapler. Methods 80 cases of inguinal hernia(hospitalized from February 2015 to January 2017)were divided into two groups according to the random number table method ,with 40 patients in each group. Traditional laparoscopic peritoneal inguinal hernia repair method was applied in the control group. Free stapler group received free stapler laparoscopic preperitoneal inguinal hernia repair treatment method. Operation time , amount of bleeding during surgery , the average hospitalization time after operation , the total cost of hospitalization,postoperative pain score,postoperative recovery activities time,patients′satisfaction,operation effusion after operation occurred scrotal hematoma and other complications were comparedbetween the two groups of patients. Results In free stapler group,patients′ satisfaction rate was significantly higher than the control group (P < 0.05). Area of effusion,scrotal hematoma and other complications infree stapler group after surgery were significantly lower than the control group (P < 0.05). Total cost of hospitalization,postoperative pain score, postoperative recovery time activities in free stapler group were significantly better than those in the control group (P<0.05). Operation time,bleeding amount during the operation,average hospitalization time difference between the two groups were not statistically significant(P > 0.05). Conclusion Operation time and amount of bleeding were similar between traditional laparoscopic transabdominal preperitoneal inguinal hernia repair method and free stapler in laparoscopic transabdominal preperitoneal inguinal hernia repair.Clinical effect of free stapler in laparoscopic transabdominal preperitoneal inguinal hernia repair proves to be effective with less complications ,less pain, faster postoperative recovery, and can reduce the cost of treatment.Free stapler in laparoscopic transabdominal preperitoneal inguinal hernia repair has satisfactory cosmetic results and was well received by patients,worthy of promotion.

Objective To investigate the epidemiology of adults acute viral gastroenteritis in Shanghai Changning district. Methods All of 1 554 stool specimens of adults acute gastroenteritis in Shanghai Changning district from June 2010 to December 2013, and the enzyme-linked immunosorbent assay and multiple polymerase chain reaction was used to detecte different viruses. Results In all of 1 554 cases, the average age was (46.19 ± 15.59) years. Among them, 691 persons were male, 863 persons were female. Virus infection was detected in 407 cases, and the detection rate was 26.19%. Among them, 395 cases (97.05%) were single virus infection, and 12 cases (2.95%) were mixed infection. The peak of epidemic was from every November to next February. The incidence of watery diarrhea, vomiting and fever in virus positive group was significantly higher than that in virus negative group:95.09%(387/407) vs. 88.14%(1 011/1 147), 31.20%(127/407) vs. 18.83%(216/1 147), and 11.06%(45/407) vs. 7.59%(87/1147), P<0.01 or<0.05. Conclusions Rotavirus infection is common in adults with acute viral gastroenteritis. Patients with positive virus infection had a higher incidence of watery diarrhea, vomiting and fever. The peak of epidemic is winter.

OBJECTIVETo investigate the effect of Kiss-1 gene suppression on the metastatic capacity of HCT116 human colorectal carcinoma cells in vitro and the involvement of nuclear factor-κB (NF-κB) signaling pathway.

METHODSA recombinant lentiviral vector of Kiss-1 gene pGC-LV-Kiss-1-EGFP or the empty vector was transfected in HCT116 cells. Cell Counting Kit-8 (CCK8) and Transwell chamber assay were used to detect the changes in cell proliferation, invasion and migration ability after the transfection. Western blotting was used to detect the expression of I-κB, the inhibitive protein of NF-κB signal pathway, and the expression of the downstream effector MMP-9 before and after transfection.

RESULTSIn cells over-expressing Kiss-1, I-κB expression increased and MMP-9 expression decreased significantly compared to those in the blank control and vector-transfected cells (P<0.05). Kiss-1 gene over-expression resulted in significant inhibition of HCT116 cell proliferation, invasion, and migration as compared to the control cells (P<0.05).

CONCLUSIONLentivirus-mediated Kiss-1 gene over-expression can inhibit the proliferation, invasion, and migration of HCT116 cells via the NF-B signaling pathway.

Cell Movement ; Cell Proliferation ; Colorectal Neoplasms ; pathology ; Genetic Vectors ; HCT116 Cells ; Humans ; I-kappa B Kinase ; metabolism ; Kisspeptins ; genetics ; Lentivirus ; Matrix Metalloproteinase 9 ; metabolism ; NF-kappa B ; metabolism ; Neoplasm Invasiveness ; Signal Transduction ; Transfection

Objective Onto investigate the indoor quality control method for qualitatively detecting the laboratory indicators of TORCH infection (rubella virus IgG ,cytomegalovirus IgG and IgM ,toxoplasma IgG and IgM ) .Methods The statistical method , normal distribution data ,ratio and standard deviation of positive rate detected by the ELISA method were adopted ,1+2s was set as the out of control rules ,the semi Lerey‐Jennings quality control chart was drawn;the direct probability calculation method was a‐dopted for the non‐normal distribution data and small probability event .The testing data of 57 batches were retrospectively ana‐lyzed .Results The positive rate of rubella virus IgG was 86 .66% ,cytomegalovirus IgG/IgM positive rates were 98 .87% and 0 .13% ,toxoplasma gondii IgG/IgM positive rates were 2 .43% and 1 .71% ,the data of 151 ,3 ,5 ,176 ,27 samples had the critical value range of five indicators .The number of out of control was once for cytomegalovirus IgG ,once and 4 times for Toxoplasma gondii IgG/IgM .Conclusion The indoor quality control for the ELISA qualitative detection of TORCH infection can adopt the data of daily detection positive rate or negative rate for monitoring the false positive .The critical value range of specimens should be fur‐ther conducted the recheck or confirmation experiment .

Objective To research the effect of the Kiss-1 gene promoter methylation on the Kiss-1 gene expression in colorectal carcinoma tissue,and to analyze the relationship between the Kiss-1 gene methylation and the clinical pathological features of colorectal carcinoma and its clinical significance.Methods The Kiss-1 gene promotor region methylation,Kiss-1 gene mRNA and protein expressions were detected respectively by methylation-specific PCR, Real-time PCR and Western blotting method in 126 cases of colorectal carcinoma tissue and para-cacinoma normal colorectal tissue.Results The positive rate of Kiss-1 gene methylation in colorectal carcinoma tissue (83.33%)was significantly higher than that in normal tissue (30.16%)(P<0.05).In the cancer tissue,the expression levels of Kiss-1 gene mRNA and protein in Kiss-1 gene promoter methylation positive group was lower than that in Kiss-1 gene promoter methylation negative group (P < 0.05 ). The positive rate of Kiss-1 gene promoter methylation in T3+T4 stage group was higher than that in T1+T2 stage group (P<0.05),but the Kiss-1 gene mRNA and protein expression levels were lower than those in T1+T2 stage group (P<0.05).In the poorly and moderately differentiated group,the positive rate of Kiss-1 gene promoter methylation was higher than that in well differentiated group (P<0.05 ), but the mRNA expression level of Kiss-1 gene was lower than that in well differentiated group (P<0.05 ). In lymph node metastasis group, the positive rate of Kiss-1 gene promoter methylation was higher than that in lymph node negative group (P<0.05),but the mRNA and protein expression levels were lower than those in lymph node negative group (P<0.05 ). And the positive rate of Kiss-1 gene promoter methylation in distant metastasis group was higher than that in non-distant metastasis group (P<0.05), but the mRNA and protein expressions levels were lower than those in non-distant metastasis group (P<0.05). There were no significant associations between the positive rate of Kiss-1 gene promoter methylation and the gender, age,tumor location, and tumor diameter of the patients (P>0.05 ). Conclusion The Kiss-1 gene promoter methylation in colorectal carcinoma tissue is associated with the Kiss-1 gene expression level and the malignant characteristics of colorectal carcinoma;Kiss-1 gene promoter methylation may be used as a reference indicator for predicting the risk of metastasis of colorectal carcinoma.